JP6462101B2 - Collagen hydrolyzate and use thereof - Google Patents

Description

  The present invention relates to a collagen hydrolyzate and to its use for the treatment and / or prevention of cellulite.

  Cellulite causes undesired changes in the properties of the skin and is apparently visible as the formation of dents on the skin surface. It is almost exclusively women who are affected, and about 80% to 98% of women are involved with aging. Cellulite occurs, among other things, in areas containing significant subcutaneous adipose tissue, namely the waist, hips, abdomen, upper thighs and upper arms.

  According to the latest research, cellulite is caused by specific changes in the dermis and subcutaneous connective tissue, especially the contraction of the septum formed from collagen fibers that connect the reticulated layer to the muscles lying under the subcutaneous fat It is believed that. This generally results in a loss of skin elasticity.

  Known therapies for cellulite include physical methods such as lymph drainage, ultrasound or vacuum, among others, but these typically do not yield success or at least do not sustain success. Since the upper skin layer of the epidermis is not directly involved in the phenomenon, topical use of cosmetics such as creams or ointments also does not allow treatment of any cause of cellulite.

  Through oral intake of collagen hydrolysates, it is known that beneficial effects can be achieved with regard to skin health in humans (V. Zague: “A new view condensing the effects of collagenate catalyst on skin propylene. ", Arch. Dermatol. Res. 2008 (9) 479). Accumulation of orally ingested collagen hydrolyzate occurs there particularly due to the ability to absorb suitable small molecule collagen peptides and good perfusion of the skin, the concentration of which is maximal in a period of about 12-24 hours after ingestion. (See M. Watanabe-Kamiyama et al .: “Absorption and effective of nominally low molecular weight hydrated hydrates.

  Against this background, the present invention proposes a new approach to the causal treatment and / or prevention of cellulite, which involves the use of collagen hydrolysates.

  Therefore, one aspect of the present invention relates to a collagen hydrolyzate as an active ingredient for treating and / or preventing cellulite.

  Clinical trials have also shown that skin elasticity increases measurable after administration of collagen hydrolyzate. This effect is particularly noticeable in women over 50 years of age. Greater skin elasticity results in severe cellulite reduction.

  Furthermore, various in vitro studies indicate that the synthesis of extracellular matrix proteins in dermal connective tissue is stimulated by collagen hydrolysates. These proteins formed by skin cells (dermal fibroblasts) include collagen (especially type I), elastin and proteoglycans (eg biglycan, versican and decorin). The synthesis of sufficient amounts of these proteins is crucial for the formation and regeneration of the extracellular matrix of the skin, which in turn is essential for dermal properties such as elasticity, resilience and moisture regulation. Is a determinant.

FIG. 1A shows a graph relating to stimulation of synthesis of type I collagen, biglycan or versican. FIG. 1B shows a graph relating to stimulation of synthesis of type I collagen, biglycan or versican. FIG. 1C shows a graph relating to stimulation of type I collagen, biglycan or versican synthesis. FIG. 2A shows a graph for increased skin moisture in hairless mice. FIG. 2B shows a graph for increased skin moisture in hairless mice. 2 shows a graph relating to stimulation of CE protein synthesis. FIG. 4A shows MALDI mass spectra of various collagen hydrolysates. FIG. 4B shows MALDI mass spectra of various collagen hydrolysates. FIG. 4C shows MALDI mass spectra of various collagen hydrolysates. FIG. 5A shows a graph for stimulation of synthesis of type I collagen, decorin and versican. FIG. 5B shows a graph for stimulation of synthesis of type I collagen, decorin and versican.

  The collagen hydrolyzate used in connection with the present invention preferably has a relatively low molecular weight. Preferably, at least 90% by weight of the collagen hydrolyzate has a molecular weight of less than 3,500 Da, more preferably at least 45% by weight has a molecular weight of less than 1,500 daltons. It has been found that a more pronounced effect can be achieved especially with low molecular weight components. The molecular weight distribution of the collagen hydrolyzate is subject to relevant limits and can be determined very accurately and reproducibly by, for example, gel permeation chromatography using a calibration standard composed of defined collagen fragments. it can.

  The average molecular weight (mass average molar mass Mw) of the collagen hydrolyzate used according to the present invention is typically in the range of about 1,700 Da to about 2,300 Da.

  In a preferred embodiment of the invention, the collagen hydrolyzate comprises at least four characteristic peptides having a molecular weight between 600 Da and 1,200 Da. Collagen hydrolysates include peptides with different chain lengths or molecular weights that occur when collagen protein chains are split, where the molecular weight distribution of these peptides is significantly different depending on the manufacturing conditions of the hydrolysates There is. Surprisingly, collagen hydrolysates having the above properties have been found to have a particularly advantageous effect on the synthesis of matrix proteins. That is, it shows significantly better results than the collagen hydrolyzate containing no characteristic peptide.

  The presence of characteristic peptides of collagen hydrolyzate can be determined in particular by MALDI mass spectrometry where the characteristic peptides appear as peaks in the mass spectrum. Preferably, at least four characteristic peptides in the molecular weight distribution found using mass spectrometry determined by MALDI have an intensity of at least 2 times, more preferably at least 4 times compared to their surroundings.

  In a preferred embodiment of the present invention, the collagen hydrolyzate comprises a peptide of 620 Da to 690 Da, a peptide of 790 Da to 860 Da, a peptide of 980 Da to 1,050 Da, and a peptide of 1,175 Da to 1,245 Da. The collagen hydrolyzate can also have a characteristic peptide of 1,500-3,500 Da.

  Preferably, the collagen hydrolyzate has a hydroxyproline content of 12% by weight or more. The amino acid hydroxyproline formed by post-translational hydroxylation of proline occurs exclusively in collagen, so that a high proportion of hydroxyproline in the collagen hydrolyzate is a widespread of other connective tissue proteins such as elastin and proteoglycans. Provide a measure of absence. These fragments can be included in specific amounts in the collagen hydrolyzate depending on the production method.

  Collagen hydrolyzate is advantageous when it is produced by enzymatic hydrolysis of gelatin. Gelatin contains denatured collagen and is obtained from connective tissues or bones of various animal species by various methods known to those skilled in the art. In the context of the present invention, the gelatin used as starting material for the collagen hydrolyzate is preferably extracted from the skin of animals, in particular pig or cow, although the use of gelatin from poultry is not excluded. Porcine gelatin, in particular pig skin gelatin, is particularly suitable as a starting material.

  Enzymatic hydrolysis of gelatin is typically performed by endoproteases to affect the amino acid profile of the resulting collagen hydrolyzate and thus increase the positive effect of the hydrolyzate It is preferred in the context of the present invention to use a plurality of endoproteases (ie at least two different endoproteases).

  According to a preferred embodiment of the invention, the collagen hydrolyzate is at least two endoproteases with different specificities, in particular at least two different metalloproteases and / or serine proteases, ie before or after certain amino acids. Are produced by the continuous action of a protease that splits the amino acid sequence of the collagen molecule. Conveniently, the metalloprotease and / or the serine protease may be the microorganisms Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciles, It is an enzyme derived from Aspergillus meleus.

  By selecting an appropriate endoprotease, not only can a collagen hydrolyzate with a characteristic molecular weight distribution be obtained, but the terminal amino acid type of the peptide obtained in the hydrolyzate will also be affected. In this context, for example, it is preferred if at least 50% of the N-terminal amino acids of the collagen hydrolyzate are hydrophobic amino acids such as the preferred alanine, leucine and isoleucine.

  According to a preferred embodiment of the invention, the collagen hydrolyzate is provided for enteral administration, particularly in the form of oral intake. Upon oral ingestion, more effective transport of collagen hydrolyzate through the blood circulation to the site of action, ie, dermal fibroblasts, is provided than with topical administration. Moreover, this dosage form is typically associated with significantly less effort for the user.

  Since the collagen hydrolyzate is extracted from raw materials permitted under the food legislation, it can be used in the context of the present invention, preferably as a dietary supplement, for treating and / or preventing cellulite. Such dietary supplements can be identified as “nutritional supplements” or “nutritional cosmetics”.

  The dietary supplement can be provided in almost any form, for example in tablets, capsules, dragees, troches, sachets or gels or solutions (eg in a single ampoule or beverage).

  Alternatively, the collagen hydrolyzate may be included in an instant powder for making food or premium food items such as confectionery or drinks. Thus, the hydrolyzate can be consumed by the user without additional effort in the context of normal nutrients (as “functional food”). In this respect, the collagen hydrolyzate is particularly advantageous when it has substantially no flavor.

  Suitably, a daily intake of about 1.5 g to 5 g, preferably about 2 g to 3 g, more preferably about 2.3 g to 2.7 g of collagen hydrolyzate is provided. Through oral intake of this amount of hydrolyzate, it has been found that significant effects that cannot be substantially increased by increasing the daily dose can be achieved.

  When used according to the present invention, the collagen hydrolyzate can be combined with other active ingredients having an advantageous effect on health, in particular on skin health, in particular with active ingredients having an antioxidant action. Such active ingredients are preferably vitamins, especially vitamins C and E, minerals, omega-3 fatty acids, omega-6 fatty acids, omega-9 fatty acids, biotin, lutein, lycopene, caffeine, glucosamine, chondroitin, hyaluron Selected from acids, folic acid, amino acids, ubiquinone 10, superoxide dismutase, and plant extracts from rosehip, lemon verbena or green tea.

  In a preferred embodiment of the present invention, administration of collagen hydrolyzate is provided to treat and / or prevent cellulite, especially in women older than 50 years, typically in postmenopausal women. In this age group, which is generally severely affected by cellulite, the effect is particularly pronounced and is demonstrated by the clinical trials described below.

  The present invention also relates to a method for treating and / or preventing cellulite in a patient, the method comprising administering a collagen hydrolyzate to the patient, particularly in the form of oral ingestion.

  Preferred embodiments of the method have already been described in connection with the use according to the invention, in particular with regard to the properties of the collagen hydrolyzate and the dose to be administered.

  The invention also relates to the use of a collagen hydrolyzate to treat and / or prevent stretch marks in the form of so-called pregnancy stretch marks (gestation line striae), particularly occurring in pregnant women. The cause of these stripes is a fine tear in the subcutaneous connective tissue due to severe stretching of the skin. Similar to cellulite, the generation of such water droplets can be offset by increasing the elasticity of the skin by administration of collagen hydrolyzate.

  A further aspect of the invention relates to the use of a collagen hydrolyzate to treat and / or prevent the occurrence of local damage to the skin as a result of pressure caused by decubitus ulcers, such as bed sores. This involves the skin receiving external pressure and its negative effects can be mitigated by increasing the elasticity of the skin.

  These and other advantages of the present invention will now be described in more detail based on the following examples and with reference to the drawings.

1． Production and Properties of Collagen Hydrolyzate To produce collagen hydrolyzate for use according to the present invention, an aqueous pig skin gelatin solution (type A) at a concentration between 20% and 40% by weight (of dry material) 200 g to 250 g of Bloom) is used as starting material. Gelatin is hydrolyzed enzymatically by continuous action of two different endoproteases of microbial origin at 50-60 ° C for 120-180 minutes, where the first enzyme is Bacillus subtilis or Bacillus An endoprotease derived from amyloliquefaciens is used, and an endoprotease derived from Bacillus licheniformis is used as the second enzyme. Subsequently, the enzyme is thermally deactivated and the solution is spray dried.

The molecular weight distribution of the obtained collagen hydrolyzate can be determined by gel permeation chromatography using the following parameters.
Static phase: TSK 2000 SW XL (Tosoh Bioscience GmbH)
Mobile phase: 0.4 mol / l sodium dihydrogen phosphate (pH 5.3)
Flow rate: 0.5 ml / min Calibration standard: Defined type I collagen fragment (FILK, Freiberg)
Detection: UV detector Knauer K-2501 (214 nm)

  As described in Table 1 below, the determination resulted in a molecular weight distribution for this collagen hydrolyzate (hereinafter referred to as the low molecular weight hydrolyzate). For comparison purposes, Table 1 shows the molecular weight distribution of commercially available collagen hydrolysates (hereinafter also referred to as polymer hydrolysates) determined using the same method.

  The hydroxyproline content of this low molecular weight hydrolyzate is about 12% to 13% by weight, which can be determined photometrically by subsequent oxidation with chloramine-T and conversion with p-dimethylaminobenzaldehyde. it can. More than 50% of the N-terminal amino acids of the hydrolyzate are hydrophobic amino acids, especially alanine, leucine and isoleucine.

2. Clinical study on the effectiveness of collagen hydrolyzate on cellulite The efficacy of the low molecular weight collagen hydrolyzate produced according to Example 1 for the treatment and / or prevention of cellulite was double-blind randomized placebo Investigated in a control study. The test subjects were 69 healthy women aged 35.3-55.4 years, divided into three groups of 23 subjects each. 68 subjects successfully completed the study.

  Starting 6 weeks before the start of the study, the use of dermatological treatment was not allowed. Subjects should also not change their life and nutrition week during the study, should not take any additional nutritional supplements and vitamin preparations, and should not expose their skin to intense UV radiation It was. Cosmetic preparations should not be used on the palmar side of the forearm, where the influence of collagen hydrolyzate on skin properties should be investigated.

Of the three groups, over a period of 8 weeks, the first group received 2.5 grams of collagen hydrolyzate daily (morning) and the second group received daily (2.5 grams each morning and afternoon). The third group received a placebo.
Daily 25 g collagen hydrolyzate, the third received placebo. The hydrolyzate could be dissolved in water or a cold drink (except milk) for ingestion.

Prior to the first intake, after 4 weeks and 8 weeks, the following skin parameters were measured on the volar side of the subject's left upper arm:
-Skin elasticity according to Cutometer (R) SEM575 (average value from three measurements)
-Transdermal water loss (TEWL) with DermaLab® device
(Average value from three measurements)
-Skin moisture content with Corneometer® CM825 (average value from 10 measurements)

  All measurements were performed 30 minutes after acclimatization in the climate control room at a temperature of 21.5 ° C. (± 1 ° C.) and a relative air humidity of 50% (± 5%).

  All three parameters were significantly increased in the group treated with collagen hydrolyzate both after 4 and 8 weeks. The measured values after 8 weeks compared to the placebo group are given in Table 2 below, especially as a percentage increase.

  Increased skin elasticity indicates the effectiveness of oral administration of collagen hydrolyzate to treat and / or prevent cellulite. Improvement of TEWL and skin moisture is a further beneficial effect of the hydrolyzate in skin health, especially resulting in increased epidermal barrier function.

  Tests for increased skin elasticity, differentiated by age group, produced the results listed in Table 3. Women under 50 years (average age 44.1 years) were compared with women over 50 years (average age 53.0 years).

  Here, what is striking is the particularly significant improvement in the skin elasticity of women over 50, thus women over 50 represent a preferred target group for use according to the invention of collagen hydrolysates.

  Skin elasticity was measured again for 4 weeks after the end of the 8 week administration period. The 92% -98% increase measured after 8 weeks is still retained, suggesting a longer lasting effect of collagen hydrolysis.

3. Stimulation of extracellular matrix protein synthesis in vitro Collagen (type I) synthesis and stimulation of proteoglycan biglycan and versican synthesis were studied in human dermal fibroblasts (skin cells) in vitro. For this purpose, the cells are incubated for 24 hours with either 0.5 mg / ml of low molecular weight or high molecular weight hydrolysates, and then expression of collagen RNA, biglycan RNA and versican RNA is determined by real-time PCR. And evaluated semi-quantitatively (compared to a control without hydrolyzate).

  The results are shown as type I collagen in FIG. 1A, biglycan in FIG. 1B, and versican in FIG. 1C, where the graph notation each represents an average value from at least 18 measurements. Shown on the horizontal axis is RNA expression relative to the control (n = 1). The left solid line column shows the control in each case, while in the middle, the shaded column is the high molecular weight hydrolyzate and the right dashed column is the low molecular weight hydrolyzate.

  Although the synthesis of all three matrix proteins is stimulated by both collagen hydrolysates, the positive effect of low molecular weight hydrolysates is more strongly expressed in each case than high molecular weight hydrolysates. Obviously. In addition to elastin, the increased effectiveness of low molecular weight hydrolysates is particularly clearly demonstrated for collagen, which is primarily responsible for skin resilience and elasticity, and for versican, which plays an important part in skin moisture regulation. ing.

  These stimulating properties of collagen hydrolysates in different matrix proteins also provide a starting point for diseases such as psoriasis, apart from the treatment and / or prevention of cellulite according to the invention, where the natural function of the skin is It is damaged.

4). Increased skin moisture content in animal studies The effect of skin moisture content using collagen hydrolysates was examined directly using hairless mice. Hairless mice are often an established model system used for dermatological investigations, and the knowledge gained from it can in principle be applied to human skin (eg, T Fujimura et al .; J. Dermatol. Sci. 2000 (24) 105-111, Y. Nishimori et al .; J. Invest. Dermatol. 2001 (117) 1458-1463).

Animals were fed daily at 150 μg collagen hydrolyzate / kg body weight for a total period of 3 weeks, while the control group received BSA instead. At the same time, all animals received a weekly UV-B irradiation dose of 18 mJ / cm ² skin, thereby negatively affecting skin moisture content.

  The water content was measured after 1 and 3 weeks using a Corneometer CM825 (manufacturer, Curage & Kazaka). The measurement principle here is based on a change in the electrostatic dose of the measuring capacitor due to the dielectric constant of the water bound to the upper skin layer, which is significantly different from the dielectric constant of most other substances.

  The results are shown as a bar chart for the measurement after 1 week in FIG. 2A and for the measurement after 3 weeks for FIG. 2B, and the notation of the graph indicates 7 measured values + standard error, respectively. What is shown on the horizontal axis is the amount of skin moisture relative to the control (n = 1). The solid line column on the left shows the control in each case, and in the middle, the shaded column is the high molecular weight hydrolyzate and the broken line column on the right is the low molecular weight hydrolyzate.

  The increase in skin moisture using the low molecular weight hydrolyzate appears to be greater both after 1 and 3 weeks than the high molecular weight hydrolyzate.

5). Stimulation of CE protein synthesis in vitro The so-called “cornified membrane” proteins play an important role in the barrier function of the skin against the invasion of pathogenic microorganisms and toxic substances. The synthesis of CE proteins, involucrin, loricrin and filaggrin, was previously determined in hairless mice fed with 150 μg collagen hydrolyzate / kg body weight (as above) daily for 5 weeks. Protein quantification relative to the control group (given BSA) was performed by SDS polyacrylamide gel electrophoresis and Western blot using specific antibodies after protein extraction from the skin.

  A result is shown as a histogram of FIG. 3, and the description of a graph shows the average value + standard deviation of seven measurements. Shown on the horizontal axis is CE protein quantification after diet using low molecular weight hydrolysates relative to controls (n = 1). The left column shows involucrin, the middle column shows loricrin, and the right column shows filaggrin.

  It is clear that the synthesis of all three CE proteins investigated is stimulated by oral intake of collagen hydrolysates, and in the case of involucrin, it is actually more than three times.

6). Analysis of molecular weight distribution using MALDI-MS A low molecular weight collagen hydrolyzate produced according to Example 1 having an average molecular weight of about 2,000 Da (hereinafter referred to as hydrolyzate A) is about 2,100 Da ( Hereinafter referred to as hydrolyzate B) and two commercially available collagen hydrolysates having an average molecular weight of about 2,900 Da (hereinafter referred to as hydrolyzate C).

The exact molecular weight distribution of these three hydrolysates was analyzed by MALDI mass spectrometry (MALDI-MS). For this purpose, samples were adjusted to a final concentration of 10 μg / μl in 0.1% trifluoroacetic acid and then purified using μC ₁₈ material. Samples were prepared using an HCCA matrix on a MALDI target and mass spectra were determined using an Ultraflex-III-TOF / TOF mass spectrometer (manufacturer: Bruker Daltonics).

  4A-4C show the corresponding mass spectra or molecular weight distributions of collagen hydrolysates A, B and C, where the molecular weight or mass number is represented on the vertical axis and the intensity is represented on the horizontal axis. A comparison of the three spectra shows that hydrolyzate A contains the following characteristic peptides, as shown in Table 4, and the relative peaks are 2 to 4 times more intense than their surroundings.

  Specifically, the four peptides of 600 Da to 1,500 Da do not correspond to the two commercially available hydrolysates B and C and are therefore particularly characteristic for hydrolysate A.

7). Stimulation of extracellular matrix protein synthesis in vitro The synthesis of collagen (type I) and proteoglycan decorin and versican was examined in vitro in human dermal fibroblasts (skin cells). For this purpose, cells are incubated for 24 hours with 0.5 mg / ml of hydrolysates A, B and C, respectively, then the expression of collagen RNA, decorin RNA and versican RNA is determined by real-time PCR, half Quantitatively evaluated. Decorin plays an important role in the formation of collagen fibers in the skin.

  The results are shown as a bar chart for hydrolyzate B in FIG. 5A and for hydrolyzate C in FIG. 5B, and the horizontal axis represents a commercially available hydrolyzate as compared to RNA expression using hydrolyzate A, respectively. Represents RNA expression in B and C (= 1). The left column represents type I collagen, the center column represents decorin, and the right column represents versican. In each case, the mean value from at least 7 measurements is shown with standard error.

  Interestingly, the data show that, with all three matrix proteins, significantly less stimulation of RNA synthesis compared to hydrolyzate A, with both slightly higher molecular weight hydrolysates B and C. Show what happens. Thus, the characteristic peptide of hydrolyzate A appears to play a critical role in its advantageous effects.

8). Exemplary recipes for nutritional (supplementary) products Several exemplary recipes for use according to the invention of collagen hydrolysates are given below, but these can usually be modified in many ways.

Capsette (dietary supplement)
Glycine 53.67% by weight
Collagen hydrolyzate 21.95% by weight
Gelatin 10.08% by weight
Guar gum 6.00% by weight
Lecithin 5.00% by weight
Citric acid 2.00% by weight
Flavor (Cassis) 0.50% by weight
Orange oil 0.50% by weight
Acesulfame K 0.30% by weight

Chocolate cocoa mass 51.0% by weight
Sucrose 22.4% by weight
Cocoa butter 16.6% by weight
Collagen hydrolyzate 10.0% by weight

Beverage Water 63.00% by weight
Aloe vera concentrate 31.00% by weight
Collagen hydrolyzate 4.00% by weight
Sucrose 1.50% by weight
Citric acid 0.26% by weight
Flavoring and coloring agents 0.24% by weight
Sucralose 0.0031% by weight

Claims (14)

  A pharmaceutical composition comprising a collagen hydrolyzate as an active ingredient for treating and / or preventing cellulite, wherein the collagen hydrolyzate has an average molecular weight in the range of 1,700 Da to 2,300 Da, Collagen hydrolyzate molecular weight distribution is less than 5% by weight collagen hydrolyzate is over 7,500 Da, 12-18% by weight is 3,500-7,500 Da, 25-31% by weight is 1, 500-3,500 Da, 40-46 wt% is 500-1,500 Da, and 5-10 wt% is less than 500 Da, the collagen hydrolyzate is an enzymatic hydrolysis of porcine skin gelatin The enzymes produced by and used for the hydrolysis are the microorganisms Bacillus subtilis, Bacillus liquor A protease from Bacillus licheniformis, Bacillus amyloliquefaciens, Aspergillus oryzae and a protease from Aspergillus or Aspergillus Said pharmaceutical composition. The pharmaceutical composition according to claim 1 , wherein at least 45% by weight of collagen hydrolyzate has a molecular weight of less than 1,500 Da. At least collagen hydrolyzate, see contains at least four peptides with a molecular weight of 600Da~1,200Da in the found molecular weight distribution by MALDI mass spectrometry, wherein the peptide compared to their surrounding The pharmaceutical composition according to claim 1 or 2, which has twice the strength . At least four peptides in the molecular weight distribution was found by MALDI mass spectrometry, as compared with those near has a strength of at least 4-fold, the pharmaceutical composition according to claim 3. The collagen hydrolyzate comprises a peptide of 620 Da to 690 Da, a peptide of 790 Da to 860 Da, a peptide of 980 to 1,050 Da, and a peptide of 1,175 Da to 1,245 Da according to any one of claims 1 to 4. The pharmaceutical composition as described. The pharmaceutical composition according to any one of claims 1 to 5 , wherein the collagen hydrolyzate has a hydroxyproline content of 12% by weight or more. At least 50% of the N-terminal amino acids of collagen hydrolyzate, alanine, leucine and isoleucine, pharmaceutical composition according to any one of claims 1-6. The pharmaceutical composition according to any one of claims 1 to 7 , wherein the collagen hydrolyzate is given for oral administration. 9. The pharmaceutical composition according to claim 8 , wherein the collagen hydrolyzate is a dietary supplement and is present in the form of tablets, capsules , dragees, troches, sachets, gels or solutions. The pharmaceutical composition according to claim 8 or 9 , wherein about 1.5 to 5 g of collagen hydrolyzate is given as a daily intake. The pharmaceutical composition according to claim 10 , wherein about 2 to 3 g of collagen hydrolyzate is given as a daily intake. The pharmaceutical composition according to claim 10 , wherein the daily intake is about 2.3 to 2.7 g of collagen hydrolyzate. Vitamins, minerals, omega-3 fatty acids, omega-6 fatty acids, omega-9 fatty acids, biotin, lutein, lycopene, caffeine, glucosamine, chondroitin, hyaluronan, folic acid, amino acids, ubiquinone-10, superoxide dismutase, and rosehip, 13. A pharmaceutical composition according to any one of claims 1 to 12 , in combination with one or more further active ingredients selected from plant extracts from lemon verbena or green tea. Collagen hydrolyzate is given for administration to postmenopausal women, the pharmaceutical composition according to any one of claims 1 to 13.